Microfibril diameter in celery collenchyma cellulose: X-ray scattering and NMR evidence

Cellulose isolated from celery collenchyma is typical of the low-crystallinity celluloses that can be isolated from primary cell-walls of higher plants, except that it is oriented with high uniformity. The diameter of the microfibrils of celery collenchyma cellulose was estimated by three separate approaches: ¹³C NMR measurement of the ratio of surface to interior chains; estimation of the dimensions of the crystalline lattice from wide angle X-ray scattering (WAXS) measurements using the Scherrer equation; and the observation that microfibrils of this form of cellulose have the unusual property of packing into an irregular array from which small angle X-ray scattering (SAXS) shows features of both form and interference functions. The interference function contributing to the SAXS pattern implied a mean microfibril centre-to-centre distance of 3.6 nm, providing an upper limit for the diameter. However modelling of the scattering pattern from an irregular array of microfibrils showed that the observed scattering curve could be matched at a range of diameters down to 2.4 nm, with the intervening space more or less sparsely occupied by hemicellulose chains. The lateral extent of the crystalline lattice normal to the 200 plane was estimated as a minimum of 2.4 nm by WAXS through the Scherrer equation, and a diameter of 2.6 nm was implied by the surface: volume ratio determined by ¹³C NMR. The WAXS and NMR measurements both depended on the assumption that the surface chains were positioned within an extension of the crystalline lattice. The reliability of this assumption is uncertain. If the surface chains deviated from the lattice, both the WAXS and the NMR data would imply larger microfibril diameters within the range consistent with the SAXS pattern. The evidence presented is therefore all consistent with microfibril diameters from about 2.4 to 3.6 nm, larger than has previously been suggested for primary-wall cellulose. Some degree of aggregation may have occurred during the isolation of the cellulose, but the larger microfibril diameters within the range proposed are a consequence of the novel interpretation of the experimental data from WAXS and NMR and are consistent with previously published data if these are similarly interpreted.     

Comparison of the characteristics of cellulose microfibril aggregates isolated from fiber and parenchyma cells of Moso bamboo (<Emphasis Type="Italic">Phyllostachys pubescens)</Emphasis>

This study examined the relationship between the functions of plant cells and the characteristics of cellulose microfibril aggregates in the cell walls. For this purpose, the mature bamboo (Phyllostachys pubescens) culm was separated into fiber and parenchyma cells, and then the morphological and physical properties of the cellulose microfibril aggregates isolated from both cells were compared. SEM observations revealed that both fiber and parenchyma cells consist of similar microfibril aggregates approximately 15–20 nm in width. Moreover, X-ray analysis and the tensile tests of the sheets prepared from the microfibril aggregates showed that the cellulose microfibrils isolated from fiber and parenchyma cells had almost the same cellulose crystallinity and longitudinal Young’s modulus in the dry state. These results suggest that all the cellulose microfibrils synthesized in the same individual exhibit the same characteristics in the dry state regardless of cell function.     

Effects of pressurized hot water extraction on the nanoscale structure of birch sawdust

Pressurized hot water extraction with a flow-through system was used to extract hemicelluloses and lignin from birch sawdust. The structure of the extraction residue was studied on various levels. Molecular mass distributions were determined with gel permeation chromatography and the crystal structure of cellulose was characterized using wide-angle X-ray scattering (WAXS). Information on the short-range order of cellulose microfibrils and on the nanoscale pore structure was obtained with small-angle X-ray scattering (SAXS), and the micrometre scale cellular morphology was imaged with X-ray microtomography. The pressurized hot water treatment was observed to increase the lateral width of cellulose crystallites, determined with WAXS, whereas a possible small decrease in the crystallinity of cellulose compared to native wood was detected. The molecular mass of cellulose remained at a relatively high level. According to the SAXS results, a tighter lateral association of cellulose microfibrils was observed in the extracted samples, which possibly led to opening of pores between bundles of microfibrils, as indicated by an increased specific surface area. A reduction in the thickness of the fibre cell walls was evidenced by X-ray microtomography.     

Cellulose microfibril orientation in onion (Allium cepa L.) epidermis studied by atomic force microscopy (AFM) and vibrational sum frequency generation (SFG) spectroscopy

Cellulose microfibril orientation in plant cell walls changes during cell expansion and development. The cellulose microfibril orientation in the abaxial epidermis of onion scales was studied by atomic force microscopy (AFM) and sum frequency generation (SFG) vibrational spectroscopy. Onion epidermal cells in all scales are elongated along the onion bulb axis. AFM images showed that cellulose microfibrils exposed at the innermost surface of the abaxial epidermis are oriented perpendicular to the bulb axis in the outer scales and more dispersed in the inner scales of onion bulb. SFG analyses can determine the orientation of cellulose microfibrils averaged over the entire thickness of the cell wall. We found that the average orientation of cellulose microfibrils inside onion abaxial epidermal cell walls as revealed by SFG is similar to the orientation observed at the innermost cell wall surface by AFM. The capability to determine the average orientation of cellulose microfibrils in intact cell walls will be useful to study how cellulose microfibril orientation is related to biomechanical properties and the growth mechanism of plant cell walls.     

Visualization of the nanoscale pattern of recently-deposited cellulose microfibrils and matrix materials in never-dried primary walls of the onion epidermis

For more than 10 years epidermal cell layers from onion scales have been used as a model system to study the relationship between cellulose orientation, cell growth and tissue mechanics. To bring such analyses to the nanoscale, we have developed a procedure for preparing epidermal peels of onion scales for atomic force microscopy to visualize the inner surface (closest to the plasma membrane) of the outer epidermal wall, with minimal disturbance and under conditions very close to the native state of the cell wall. The oriented, multilayer distribution of cellulose microfibrils, approximately ~3 nm wide, is readily observed over extended lengths, along with other features such as the distribution of matrix substances between and on top of microfibrils. The microfibril orientation and alignment appear more dispersed in younger scales compared with older scales, consistent with reported values for mechanical and growth anisotropy of whole epidermal sheets. These results open the door to future work to relate cell wall structure at the nm scale with larger-scale tissue properties such as growth and mechanical behaviors and the action of cell wall loosening agents to induce creep of primary cell walls.     

Structure and spacing of cellulose microfibrils in woody cell walls of dicots

The structure of cellulose microfibrils in situ in wood from the dicotyledonous (hardwood) species cherry and birch, and the vascular tissue from sunflower stems, was examined by wide-angle X-ray and neutron scattering (WAXS and WANS) and small-angle neutron scattering (SANS). Deuteration of accessible cellulose chains followed by WANS showed that these chains were packed at similar spacings to crystalline cellulose, consistent with their inclusion in the microfibril dimensions and with a location at the surface of the microfibrils. Using the Scherrer equation and correcting for considerable lateral disorder, the microfibril dimensions of cherry, birch and sunflower microfibrils perpendicular to the [200] crystal plane were estimated as 3.0, 3.4 and 3.3 nm respectively. The lateral dimensions in other directions were more difficult to correct for disorder but appeared to be 3 nm or less. However for cherry and sunflower, the microfibril spacing estimated by SANS was about 4 nm and was insensitive to the presence of moisture. If the microfibril width was 3 nm as estimated by WAXS, the SANS spacing suggests that a non-cellulosic polymer segment might in places separate the aggregated cellulose microfibrils.     

Vibrational sum-frequency-generation (SFG) spectroscopy study of the structural assembly of cellulose microfibrils in reaction woods

The cellulose microfibril assemblies in secondary cell walls of tension wood and compression wood were studied with vibrational sum frequency generation (SFG) spectroscopy. The tension wood contains the gelatinous layer with highly-crystalline and highly-aligned cellulose microfibrils. The SFG spectral features of tension wood changed depending on the azimuth angle between the polarization of the incident IR beam and the preferential alignment axis of the cellulose microfibrils. The SFG spectra of the compression wood did not show any dependence on the azimuth angle, implying that the overall orientation of cellulose microfibrils in compression wood is not highly aligned. Instead, the decrease of cellulose content in compression wood brought about larger separation between cellulose microfibrils, which was manifested as changes in CH₂/OH intensity ratio in SFG spectra. These results implied that SFG spectral features are sensitive to cellulose microfibril alignments and inter-fibrillar separations.     

The influence of thermo-hygro-mechanical treatment on the micro- and nanoscale architecture of wood cell walls using small- and wide-angle X-ray scattering

Tracking the changes of cellulose crystallites upon thermo-hygro-mechanical treatment is essential to understand the response of wood cell walls to steam and compression. In this paper the influence of Compression combined with Steam (CS) treatment on wood cellulose crystallites and pores structure of Chinese fir (Cunninghamia lanceolata) was studied under different steaming temperatures and compression ratios. Small-angle X-ray scattering and wide-angle X-ray scattering were used to investigate the changes of cellulose crystallites dimension, aspect ratio, fibril diameter distribution, non-crystalline fraction, the number of chains in each microfibril, as well as the fractal dimension and size of pores in response to CS treatment conditions. Results indicate that the crystallinity increased due to CS treatment, but did not show alteration with varying CS treatment conditions, i.e. seemed nearly unaffected by higher temperatures or compression ratio, both for earlywood and latewood. The cellulose crystallite diameter depended on processing parameters: it increased with increasing treatment temperature. No considerable differences were found for earlywood and latewood. We interpret our findings as a rearrangement of adjacent cellulose chains towards higher crystalline perfection attributing to the increase in crystallinity. The same effect allows a larger coherence length of crystalline order and therefore features an increasing cross-sectional dimension. In general we can state that the CS treatment leads to higher crystallinity and more perfectly arranged cellulose crystals, while it does not greatly affect the microfibril diameter but rather the amorphous regions of the microfibrils and the surrounding hemicellulose and lignin.     

Comparison of the characteristics of cellulose microfibril aggregates of wood, rice straw and potato tuber

The focus of this study has been to isolate cellulose microfibril aggregates by the one-time grinding treatment from wood, rice straw and potato tuber, and to compare their morphological and mechanical properties. Field emission scanning electron microscopy images showed that the diameter range of isolated microfibril aggregates from wood, 12–20 nm, was smaller than those from rice straw and potato tuber, 12–35 nm and 12–55 nm, respectively. These differences were observed even in the purified rice straws and potato tuber before the grinder treatment, but were hardly observed in the purified wood. The results of X-ray analysis and tensile tests indicated that there were no significant differences among the sources in the cellulose crystallinity and Young’s modulus of the isolated microfibril aggregates in the dry state. These results suggest that the inherent characteristics of cellulose microfibril aggregates in the dry state are very similar regardless of plant sources and tissue functions.     

Domain structure of nylon 6 fibers

Stained and unstained sections of nylon 6 fibers are examined by means of transmission electron microscopy. Data are presented regarding dimensions and shape of macrofibrils, microfibrils, amorphous, and crystalline domains of the microfibril and the spacing between the microfibrils. The new results support the conclusions of a previous SAXS and diffusion study carried out with the same fibers.     

Mesoporous structures in never-dried softwood cellulose fibers investigated by nitrogen adsorption

Nitrogen adsorption was used to characterize mesoporous structures in never-dried softwood cellulose fibers. Distinct inflections in desorption isotherms were observed over the relative vapor pressure (P/P₀) range of 0.5–0.42 for never-dried cellulose fibers and partially delignified softwood powders. The reduction in N₂ adsorption volume was attributed to cavitation of condensed N₂ present in mesopores formed via lignin removal from wood cell walls during delignification. The specific surface areas of significantly delignified softwood powders were ~150 m² g^?1, indicating that in wood cell walls 16 individual cellulose microfibrils, each 3–4 nm in width, form one cellulose fibril bundle surrounded with a thin layer of lignin and hemicelluloses. Analysis of N₂ adsorption isotherms indicates that mesopores in the softwood cellulose fibers and partially delignified softwood powders had peaks ranging from 4 to 20 nm in diameter.     

Bacterial cellulose–silica organic–inorganic hybrids

Bacterial cellulose (BC) hydrated membranes present nanometric reticulated structure that can be used as a template in the preparation of new organic–inorganic hybrids. BC–silica hybrids were prepared from BC membranes and tetraethoxysilane, (TEOS) at neutral pH conditions at room temperature. Macroscopically homogeneous membranes were obtained containing up to 66 wt.% of silica spheres, 20–30 nm diameter. Scanning electron micrographs clearly show the silica spheres attached to cellulose microfibrils. By removing the cellulose, the silica spheres can be easily recovered. The new hybrids are stable up to 300 °C and display a broad emission band under UV excitation assigned to oxygen-related defects at the silica particles surface. Emission color can be tuned by changing the excitation wavelength.     

Structure of cellulose and microcrystalline cellulose from various wood species, cotton and flax studied by X-ray scattering

The structure of microcrystalline cellulose (MCC) made by mild acid hydrolysis from cotton linter, flax fibres and sulphite or kraft cooked wood pulp was studied and compared with the structure of the starting materials. Crystallinities and the length and the width of the cellulose crystallites were determined by wide-angle X-ray scattering and the packing and the cross-sectional shape of the microfibrils were determined by small-angle X-ray scattering. The morphological differences were studied by scanning electron microscopy. A model for the changes in microfibrillar structure between native materials, pulp and MCC samples was proposed. The results indicated that from softwood or hardwood pulp, flax cellulose and cotton linter MCC with very similar nanostructures were obtained with small changes in reaction conditions. The crystallinity of MCC samples was 54–65%. The width and the length of the cellulose crystallites increased when MCC was made. For example, between cotton and cotton MCC the width increased from 7.1 nm to 8.8 nm and the length increased from 17.7 nm to 30.4 nm. However, the longest crystallites were found in native spruce wood (35–36 nm).     

AFM observation of ultrathin microfibrils in fruit tissues

Polysaccharide microfibrils in fruit tissue of higher plants (strawberry, peach, and rambutan) were examined by atomic force microscopy (AFM). For preserving native cellulose microfibrils, a combination of 2% NaClO₂ and 4% NaOH extraction was applied to the materials. This corresponds to mild alkali extractin of holocellulose, and denoted here as weak alkali-resistant polysaccharide, WARP. The amount of neutral sugars corresponded to 53–95% of that of WARP. Glucose was the most abundant in the neutral sugars, being 74–87%. AFM examination of microfibrils dispersed on mica substrate allowed accurate determination of fibril widths as 1.0–2.0 nm. Their straight fibrillar shape and the results of X-ray diffraction and infrared spectroscopy indicate that these fibrils are cellulose. These results constitute direct evidence for existence of ultrafine cellulose microfibrils hitherto assumed from X-ray diffraction and NMR analyses.     

Deswelling of microfibril bundles in drying wood studied by small-angle neutron scattering and molecular dynamics

Cellulose - Structural changes of cellulose microfibrils and microfibril bundles in unmodified spruce cell wall due to drying in air were investigated using time-resolved small-angle neutron...     

On the nanostructure of micrometer-sized cellulose beads

The analysis of the porosity of materials is an important and challenging field in analytical chemistry. The gas adsorption and mercury intrusion methods are the most established techniques for quantification of specific surface areas, but unfortunately, dry materials are mandatory for their applicability. All porous materials that contain water and other solvents in their functional state must be dried before analysis. In this process, care has to be taken since the removal of solvent bears the risk of an incalculable alteration of the pore structure, especially for soft materials. In the present paper, we report on the use of small-angle X-ray scattering (SAXS) as an alternative analysis method for the investigation of the micro and mesopores within cellulose beads in their native, i.e., water-swollen state; in this context, they represent a typical soft material. We show that even gentle removal of the bound water reduces the specific surface area dramatically from 161 to 109 m² g⁻¹ in cellulose bead sample type MT50 and from 417 to 220 m² g⁻¹ in MT100. Simulation of the SAXS curves with a bimodal pore size distribution model reveals that the smallest pores with radii up to 10 nm are greatly affected by drying, whereas pores with sizes in the range of 10 to 70 nm are barely affected. The SAXS results were compared with Brunauer–Emmett–Teller results from nitrogen sorption measurements and with mercury intrusion experiments.     

Atomic force microscopy and transmission electron microscopy of cellulose from Micrasterias denticulata; evidence for a chiral helical microfibril twist

Atomic force microscopy (AFM), tapping mode atomic force microscopy (TM-AFM) and transmission electron microscopy (TEM) have been used to image the cell wall, ultrathin sections of whole cells and cellulose microfibrils prepared from the green alga Micrasterias denticulata. Measurements of the microfibril dimensions are in agreement with earlier observations carried out by electron microscopy. Images at the molecular level of the surface of the microfibrils were obtained with AFM and show regular periodicities along the microfibril axis that correspond to the fibre and glucose repeat distances of cellulose. Twisted regions visible at intervals along the microfibrils dried down onto substrates were noted to be right-handed in over 100 observations by TEM, AFM and TM-AFM. This revised version was published online in August 2006 with corrections to the Cover Date.     

Multiscale modeling of the elastic properties of natural fibers based on a generalized method of cells and laminate analogy approach

In this article, a new method based on a generalized method of cells and laminate analogy approach was used to predict the elastic properties of natural fibers. The elastic properties of cellulose crystals and amorphous cellulose were adopted to calculate the effective properties of microfibrils. A ten-layer antisymmetrical laminated structure was used to predict the effective properties of cell walls. The effects of the aspect ratio and volume fraction of cellulose crystal, the microfibril angle in the S2 layer and the lumen ratio of fiber on the axial Young’s moduli of natural fibers were analyzed in detail. The results show that the predicted properties of fibers are those of the cell fibers, and the final elastic properties of natural fibers can be obtained with the volume fractions of cell fibers as the corresponding conversion coefficients. The multiscale method is very effective in the predictions of the axial Young’s moduli of natural fibers.     

Preparation and characterization of cellulose nanofibers from partly mercerized cotton by mixed acid hydrolysis

Cellulose nanofibers with a diameter of 70 nm and lengths of approximately 400 nm were fabricated from partly mercerized cotton fibers by acid hydrolysis. Morphological evolution of the hydrolyzed cotton fibers was investigated by powder X-ray diffraction, Fourier transform infrared analysis and field emission scanning electron microscopy. The XRD results show that the cellulose I was partially transformed into cellulose II by treatment with 15 % NaOH at 150° for 3 h. The crystallinity of this partially mercerized sample was lower than the samples that were converted completely to cellulose II by higher concentrations of NaOH. The intensities of all of the diffraction peaks were noticeably increased with increased hydrolysis time. Fourier transform infrared results revealed that the chemical composition of the remaining nanofibers of cellulose I and II had no observable change after acidic hydrolysis, and there was no difference between the hydrolysis rates for cellulose I or II. The formation of cellulose nanofibers involves three stages: net-like microfibril formation, then short microfibrils and finally nanofibers.     

Effect of alkaline treatment on cellulose supramolecular structure studied with combined confocal Raman spectroscopy and atomic force microscopy

The aim of this work was to investigate the morphological and structural changes associated with mercerization of cellulose fibres with combined confocal Raman and atomic force microscopy (AFM). During mercerization the alkali induces a change in polymorphic lattice from cellulose I to II. This was observed by confocal Raman spectroscopy from cellulose samples treated with 10, 15 and 25% aqueous sodium hydroxide solution. AFM images from the same samples illustrated that microfibrils were swollen and more granular in cellulose II than in cellulose I. Raman spectral images in plane and depth directions showed that the polymorphous cellulose structure was uniform throughout the cell wall, whereas the microfibril orientation varied between fibre cell wall layers. The changes in microfibril orientation on the sample surfaces were confirmed by AFM images measured from the same sample position.